We have sequenced a part of the insert of a lambda clone (EMBL3) after
subcloning in pUC19. Now we are trying to sequence into the flanking
sequences using a primer based on the sequence already known.
In order to avoid subcloning, we are looking for a protocol suitable for
sequencing lambda DNA directly. We have tried some denaturation protocols and
varied the amounts of DNA, but with little success. The sequencing primers
work well when using them on pUC-cloned DNA.
Could somebody provide a protocol suitable for the A.L.F. sequencer (using
fluorescently-labeled primers)?
Rene De Mot